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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-8855 | |||||||||
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Title | Cryo-EM structure of P. aeruginosa flagellar filaments A443V | |||||||||
![]() | Cryo-EM structure of P. aeruginosa flagellar filaments A443V | |||||||||
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![]() | bacteria flagella / helical polymers / cryo-EM / PROTEIN FIBRIL | |||||||||
Function / homology | ![]() bacterial-type flagellum / bacterial-type flagellum-dependent cell motility / structural molecule activity / extracellular region Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | helical reconstruction / cryo EM / negative staining / Resolution: 4.2 Å | |||||||||
![]() | Wang F / Postel S | |||||||||
Funding support | ![]()
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![]() | ![]() Title: A structural model of flagellar filament switching across multiple bacterial species. Authors: Fengbin Wang / Andrew M Burrage / Sandra Postel / Reece E Clark / Albina Orlova / Eric J Sundberg / Daniel B Kearns / Edward H Egelman / ![]() Abstract: The bacterial flagellar filament has long been studied to understand how a polymer composed of a single protein can switch between different supercoiled states with high cooperativity. Here we ...The bacterial flagellar filament has long been studied to understand how a polymer composed of a single protein can switch between different supercoiled states with high cooperativity. Here we present near-atomic resolution cryo-EM structures for flagellar filaments from both Gram-positive Bacillus subtilis and Gram-negative Pseudomonas aeruginosa. Seven mutant flagellar filaments in B. subtilis and two in P. aeruginosa capture two different states of the filament. These reliable atomic models of both states reveal conserved molecular interactions in the interior of the filament among B. subtilis, P. aeruginosa and Salmonella enterica. Using the detailed information about the molecular interactions in two filament states, we successfully predict point mutations that shift the equilibrium between those two states. Further, we observe the dimerization of P. aeruginosa outer domains without any perturbation of the conserved interior of the filament. Our results give new insights into how the flagellin sequence has been "tuned" over evolution.Bacterial flagellar filaments are composed almost entirely of a single protein-flagellin-which can switch between different supercoiled states in a highly cooperative manner. Here the authors present near-atomic resolution cryo-EM structures of nine flagellar filaments, and begin to shed light on the molecular basis of filament switching. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 42.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 13.2 KB 13.2 KB | Display Display | ![]() |
Images | ![]() | 101.2 KB | ||
Filedesc metadata | ![]() | 5.7 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5wk5MC ![]() 8847C ![]() 8848C ![]() 8849C ![]() 8850C ![]() 8851C ![]() 8852C ![]() 8853C ![]() 8856C ![]() 5wjtC ![]() 5wjuC ![]() 5wjvC ![]() 5wjwC ![]() 5wjxC ![]() 5wjyC ![]() 5wjzC ![]() 5wk6C C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Cryo-EM structure of P. aeruginosa flagellar filaments A443V | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.05 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Pseudomonas aeruginosa flagella filament
Entire | Name: Pseudomonas aeruginosa flagella filament |
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Components |
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-Supramolecule #1: Pseudomonas aeruginosa flagella filament
Supramolecule | Name: Pseudomonas aeruginosa flagella filament / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: B-type flagellin
Macromolecule | Name: B-type flagellin / type: protein_or_peptide / ID: 1 / Number of copies: 41 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() Strain: ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1 |
Molecular weight | Theoretical: 49.302906 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MALTVNTNIA SLNTQRNLNA SSNDLNTSLQ RLTTGYRINS AKDDAAGLQI SNRLSNQISG LNVATRNAND GISLAQTAEG ALQQSTNIL QRIRDLALQS ANGSNSDADR AALQKEVAAQ QAELTRISDT TTFGGRKLLD GSFGTTSFQV GSNAYETIDI S LQNASASA ...String: MALTVNTNIA SLNTQRNLNA SSNDLNTSLQ RLTTGYRINS AKDDAAGLQI SNRLSNQISG LNVATRNAND GISLAQTAEG ALQQSTNIL QRIRDLALQS ANGSNSDADR AALQKEVAAQ QAELTRISDT TTFGGRKLLD GSFGTTSFQV GSNAYETIDI S LQNASASA IGSYQVGSNG AGTVASVAGT ATASGIASGT VNLVGGGQVK NIAIAAGDSA KAIAEKMDGA IPNLSARART VF TADVSGV TGGSLNFDVT VGSNTVSLAG VTSTQDLADQ LNSNSSKLGI TASINDKGVL TITSATGENV KFGAQTGTAT AGQ VAVKVQ GSDGKFEAAA KNVVAAGTAA TTTIVTGYVQ LNSPTAYSVS GTGTQASQVF GNASAAQKSS VASVDISTAD GAQN AIAVV DNALAAIDAQ RADLGAVQNR FKNTIDNLTN ISENATNVRS RIKDTDFAAE TAALSKNQVL QQAGTAILAQ ANQLP QAVL SLLR UniProtKB: B-type flagellin |
-Experimental details
-Structure determination
Method | negative staining, cryo EM |
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![]() | helical reconstruction |
Aggregation state | filament |
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Sample preparation
Concentration | 0.1 mg/mL |
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Buffer | pH: 7.4 / Details: PBS |
Staining | Type: NEGATIVE / Material: negative stain |
Grid | Pretreatment - Type: PLASMA CLEANING |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Average exposure time: 2.0 sec. / Average electron dose: 20.0 e/Å2 Details: Images were stored containing seven parts, where each part represented a set of frames corresponding to a dose of ~20 electrons per Angstrom^2. The full dose image stack was used for the ...Details: Images were stored containing seven parts, where each part represented a set of frames corresponding to a dose of ~20 electrons per Angstrom^2. The full dose image stack was used for the estimation of the CTF as well as for boxing filaments. Only the first two parts were used for the reconstruction (~5 electrons per Angstrom^2). |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Final reconstruction | Applied symmetry - Helical parameters - Δz: 4.61 Å Applied symmetry - Helical parameters - Δ&Phi: 65.75 ° Applied symmetry - Helical parameters - Axial symmetry: C1 (asymmetric) Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 4.2 Å / Resolution method: OTHER / Software - Name: SPIDER / Details: model-map FSC 0.38 cut-off / Number images used: 102119 |
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Startup model | Type of model: OTHER / Details: featureless cylinder |
Final angle assignment | Type: NOT APPLICABLE / Software - Name: SPIDER |
-Atomic model buiding 1
Refinement | Space: REAL |
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Output model | ![]() PDB-5wk5: |